Source: UNIVERSITY OF TENNESSEE submitted to
ENTERIC DISEASES OF SWINE AND CATTLE: PREVENTION, CONTROL AND FOOD SAFETY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0225049
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
NC-1041
Project Start Date
Jan 1, 2011
Project End Date
Sep 30, 2012
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF TENNESSEE
2621 MORGAN CIR
KNOXVILLE,TN 37996-4540
Performing Department
Animal Science
Non Technical Summary
The estimated cases of campylobacteriosis in the United States are more than 2 million per year. The medical and productivity costs resulting from C. jejuni infection are estimated at $1.5 to $8.0 billion dollars each year in the United States. Epidemiologic studies have revealed that poultry are the major reservoir of C. jejuni, and consumption of undercooked chickens or food contaminated by poultry products are responsible for the majority of human Campylobacter enteritis. Increasing evidence also indicates that antibiotic use in poultry selects for resistant strains of C. jejuni, posing a significant threat to public health. Thus, on-farm control of C. jejuni is urgently needed to reduce human exposure and the number of foodborne illness in the U.S. Specifically, this project has following expected impacts. Impact 1. Campylobacter jejuni, the most prevalent foodborne human pathogen, is the leading bacterial cause of human gastroenteritis in the U.S. Research in this lab focuses on the development of innovative strategies to control Campylobacter infection in humans and in animal reservoirs, consequently reducing the occurrence of foodborne illness. Impact 2. Emergence of bacterial antibiotic resistance has become a serious problem worldwide. Research in this lab will elucidate the mechanisms for the development, persistence, and transfer of antibiotic resistance in zoonotic foodborne human pathogens. The studies may open new avenues for treatment and prevention of resistant foodborne pathogens important in animal health and food safety. Impact 3. Zoonotic human pathogens (e.g. Campylobacter) could be used as biological agents in acts of terrorism. Thus, research in this lab is also significant in biodefense. The studies will provide important information to develop effective vaccines, diagnostics and therapeutics to protect the public health. Impact 4. Research on the effects of antibiotic growth promoter on intestinal microbiota will lead to development of alternative microbiota-based strategies for promoting growth of food animals, which has significant impact on the production, sustainability, and health of food animals.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3113310117010%
7223220110050%
7123210104040%
Knowledge Area
311 - Animal Diseases; 722 - Zoonotic Diseases and Parasites Affecting Humans; 712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
3220 - Meat-type chicken, live animal; 3210 - Egg-type chicken, live animal; 3310 - Beef cattle, live animal;

Field Of Science
1040 - Molecular biology; 1100 - Bacteriology; 1170 - Epidemiology;
Goals / Objectives
My overall research goal is to study molecular mechanisms of bacterial pathogenesis and drug resistance, which will reveal potential targets for development of novel intervention strategies and diagnostic tools against pathogens important in animal health and food safety/security. Objectives: 1. Focus on emerging diseases- Identify, characterize and develop improved detection methods related to newly recognized, novel or emerging causes of zoonotic enteric diseases and enteric pathogens of cattle and swine. 2. Focus on effective interventions- Develop and improve interventions and preventative measures to reduce the incidence and prevalence of infections of cattle and swine with enteric and food borne disease agents. The specific objectives are: 1) To study high-affinity siderophore-mediated iron scavenging for Campylobacter infection. 2) To determine molecular basis of antimicrobial peptide reisstance in Campylobacter 3) To elucidate antibiotic resistance gene reservoir in gut microbiota and corresponding mechanisms and transmission of antimicrobial resistance. 4) To examine effect of antibiotic growth promoters on intestinal microbiota in food animals.
Project Methods
Both fundamental and contemporary approaches will be used in this project to study Campylobacter jejuni, the leading foodborne human pathogen causing enteritis in the United States and many other industrialized countries. Specifically, we will use a panel of genetic, genomics and bioinformatics approaches to achieve our goal, such as site-directed mutagenesis, random transposon mutagenesis, microarray, transformation, conjugation, quantitative Real-Time RT-PCR, Phylochips, and pyrosequencing. A powerful chicken model will be for studying mechanisms of pathogenesis and antibiotic resistance in Campylobacter and also will be used for developing innovative vaccines against Campylobacter.

Progress 01/01/11 to 09/30/12

Outputs
OUTPUTS: 1. Campylobacter a) Ferric enterobactin acquisition systems in Campylobacter. Our study identified and characterized a novel periplasmic trilactone esterase and suggested a new model of FeEnt acquisition in Campylobacter. We also firmly established that Campylobacter could utilize high-affinity salmochelin for iron acquisition, and provided insights into the delicate interaction between Campylobacter and host during infection. b) Development and evaluation of Campylobacter vaccine. In this study, we have successfully developed CfrA- and CmeC-based subunit vaccines, which provides us a solid foundation to evaluate different vaccination regimens for effective mitigation of Campylobacter in poultry in the future. c) Antimicrobial peptide resistance in Campylobacter. This study identified four genetic loci in Campylobacter that will be useful for characterizing molecular basis of Campylobacter resistance to AMPs, a significant knowledge gap in Campylobacter pathogenesis. d) Regulation of beta-lactam resistance in Campylobacter jejuni. We have identified a novel mechanism of beta-lactam resistance in C. jejuni, which will help us better understand the development and regulation of β-lactam resistance, a significant issue in many bacterial pathogens. 2. Gut microbiota: role in antibiotic resistance and animal health a) Functional cloning and characterization of antibiotic resistance genes from chicken microbiome. This study showed the effectiveness of functional cloning approach for examination of AR reservoir in food animals, revealed novel AR resistance genes in gut microbiome of chickens from different production systems, and demonstrated functional compatibility of the novel AR genes in enteric pathogens. b) Identification and Characterization of a Bile Salt Hydrolase from Lactobacillus salivarius for Developing Novel Alternatives to Antibiotic Growth Promoters. Tthis study identified and characterized a BSH with broad substrate specificity from a chicken L. salivarius strain and established a solid platform for us to discover novel BSH inhibitors, the promising feed additives to replace AGPs for enhancing the productivity and sustainability of food animals. PARTICIPANTS: Following individuals are actively involved this project during the reporting period: 1. Ximin Zeng, Postdoc 2. Yiming Mo, Postdoc 3. Barbara Gillespie, Research Associate 4. Katie Smith, MS student 5. Lindsay Jones, MS student 6. Samantha Brown, Undergraduate Research Assistant 7. Elisabeth M. Spratt, Undergraduate Research Assistant 8. Jennifer M. Hime, Undergraduate Research Assistant TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
C. jejun is the leading foodbome human pathogen in the United States and many other industrialized countries. Increasing evidence also indicates that antibiotic use in poultry selects for resistant C. jejuni, posing a significant threat to public health. We have made significant progress to understand the molecular mechanisms of pathogenesis and antibiotic resistance in Campylobacter during the reporting period. The findings have filled a significant gap in antimicrobial resistance development in Campylobacter and provided important information for the development of effective vaccine to control Campylobacter. We also made significant progress on the development of innovative alternatives to antibiotic growth promoters. Financial support from experiment station for faculty, technician and graduate student is essential for producing project outputs and achieving project outcomes and impacts. In summary, the studies have following three significant impacts: 1) The studies focus on the development of innovative strategies to control Campylobacter infection in humans and in animal reservoirs, consequently reducing the occurrence of foodborne illness. 2) Emergence of bacterial antibiotic resistance has become a serious problem worldwide. Our studies may open new avenues for treatment and prevention of resistant foodborne pathogens important in animal health and food safety. 3) Zoonotic human pathogens (e.g. Campylobacter) could be used as biological agents in acts of terrorism. Thus, our findings are also significant in biodefense. The studies will provide important information to develop effective vaccines, diagnostics and therapeutics to protect the public health.

Publications

  • Referreed Journal Articles 1. Wang, Z., X. Zeng, Y. Mo, K. Smith, J. Lin. 2012. Identification and characterization of a bile salt hydrolase for developing novel alternatives to antibiotic growth promoters. Applied and Environmental Microbiology 78:8795-8802
  • 2. Sanad, Y.M., I.I. Kassem, Z. Liu, J. Lin, J. T. Lejune, and G. Rajashekara. 2012. Occurrence of the invasion associated marker (iam) in Campylobacter jejuni isolated from cattle. BMC Research Notes. 4:570
  • Scientific Abstracts 1. Zeng, X., B. Gillespie, S. Brown, J. Lin. 2012. Cj0843c, a putative lytic transglycosylase, is involved in beta-lactam resistance by modulating beta-lactamse activity in Campylobacter jejuni. 93rd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 2-4, Chicago
  • 2. Smith, K., Z. Wang, X. Zeng, Y. Mo, J. Lin. 2012. Discovery of novel alternatives to antibiotic growth promoter to protect food safety. 93rd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 2-4, Chicago
  • 3. Mo, Y., X. Zeng, J. Lin. 2012. Salmochelin-mediated iron acquisition in Campylobacter jejuni. 93rd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 2-4, Chicago.
  • 4. Jones, L., X. Zeng, and J. Lin. 2012. Development of novel vaccines for mitigation of Campylobacter in poultry. 93rd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 2-4, Chicago
  • 5. Lin, J. W. Zhong, X. Zeng, Y. Mo, K. Smith. 2012. Identification of bile salt hydrolase inhibitors, the promising alternative to antibiotic growth promoters. International Symposium of Alternatives to Antibiotics: Challenges and Solutions in Animal Production. September 25-28, Paris, France.
  • 6. Lin, J., W. Zhong, X. Zeng, Y. Mo, K. Smith. 2012. High throughput genome sequencing identified a bile salt hydrolase for developing novel alternatives to antibiotic growth promoters. Invited for "Next Generation Sequencing Tools: Applications for Food Safety and Poultry Production" Symposium in 2012 Poultry Science Association Annual Meeting, July 9-12, Athens, Georgia.
  • 7. Zeng, X., F. Xu, and J. Lin. 2012. Molecular interactions of high-affinity ferric enterobactin acquisition systems in Campylobacter. 112th General Meeting of American Society for Microbiology, June 16-19, San Francisco
  • 8. Xu, F., X. Zeng, B. Yang, A. Shi, J. Lin. 2012. Role of Ferric Enterobactin Receptors in Norepinephrine-mediated Growth Promotion in Campylobacter. 112th General Meeting of American Society for Microbiology, June 16-19, San Francisco
  • 9. Mo, Y., X. Zeng, and J. Lin. 2012. In vitro characterization of Cee, a novel periplasmic ferric enterobactin esterase in Campylobacter. 112th General Meeting of American Society for Microbiology, June 16-19, San Francisco
  • 10. Wang, Z., K. Smith, X. Zeng, Y. Mo, and J. Lin. 2012. Identification and characterization of a bile salt hydrolase for developing novel alternatives to antibiotic growth promoters. 112th General Meeting of American Society for Microbiology, June 16-19, San Francisco
  • 11. Smith, K., Z. Wang., X. Zeng, Y. Mo, and J. Lin. 2012. Identification of bile salt hydrolase inhibitors: the promising alternative to antibiotic growth promoters. Comparative & Experimental Medicine and Public Health Research Symposium. University of Tennessee, May 21-22, Knoxville, TN
  • 12. Mo, Y., X. Zeng, and J. Lin. 2012. In vitro characterization of Cee, a novel periplasmic ferric enterobactin esterase in Campylobacter. Comparative & Experimental Medicine and Public Health Research Symposium. University of Tennessee, May 21-22, Knoxville, TN
  • 13. Zeng, X., B. Gillespie, S. Brown, J. Lin. 2012. Identification of Cj0843c, a putative lytic transglycosylase involved in beta-lactam resistance in Campylobacter jejuni. Comparative & Experimental Medicine and Public Health Research Symposium. University of Tennessee, May 21-22, Knoxville, TN


Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: The findings from this project have been disseminated to scientific communities mainly in two ways: 1. Presentation in national and international conferences. The results were presented at 1) 16th International Workshop on Campylobacter, Helicobacter and Related Organisms. Aug 28 - Sep 1, Vancouver, Canada; 2) Fourth International Rushmore Conference on Enteric Diseases. Dec 3-4, Chicago; and 3) 92nd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 4-6, Chicago. 2. Publication in peer-reviewed scientific journals. The results have been published in high impact journals such as Journal of Applied Microbiology, Frontiers in Cellular and Infection Microbiology, Applied and Environmental Microbiology. PARTICIPANTS: Ximin Zeng, Postdoc Yiming Mo, Postdoc Wei Zhou, MS Student Katie Smith, MS Student Samantha Brown, Undergraduate Research Assistant Jennifer M Hime, Undergraduate Research Assistant TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
C. jejuni is the leading foodbome human pathogen in the United States and many other industrialized countries. Increasing evidence also indicates that antibiotic use in poultry selects for resistant C. jejuni, posing a significant threat to public health. We have made significant progress to understand the molecular mechanisms of pathogenesis and antibiotic resistance in Campylobacter during the reporting period. The findings have filled a significant gap in antimicrobial resistance development in Campylobacter and provided important information for the development of effective vaccine to control Campylobacter. Financial support from experiment station for faculty, technician and graduate student is essential for producing project outputs and achieving project outcomes and impacts. In summary, the studies have following three significant impacts: 1. The studies focus on the development of innovative strategies to control Campylobacter infection in humans and in animal reservoirs, consequently reducing the occurrence of foodborne illness. 2. Emergence of bacterial antibiotic resistance has become a serious problem worldwide. Our studies may open new avenues for treatment and prevention of resistant foodborne pathogens important in animal health and food safety. 3. Zoonotic human pathogens (e.g. Campylobacter) could be used as biological agents in acts of terrorism. Thus, our findings are also significant in biodefense. The studies will provide important information to develop effective vaccines, diagnostics and therapeutics to protect the public health.

Publications

  • Zeng, X., F. Xu, Y. Mo., and J. Lin. 2011. Characterization of enterobactin esterase Cee in Campylobacter led to a novel model of ferric enterobactin iron acquisition. 92nd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 4-6, Chicago
  • Hoang, K.V., N. J. Stern, and J. Lin. 2011. Development and stability of bacteriocin resistance in Campylobacter. Journal of Applied Microbiology.111:1544-1550.
  • Lin, J. 2011. Effect of antibiotic growth promoters on intestinal microbiota in food animals: a novel model for studying the relationship between gut microbiota and human obesity Frontiers in Cellular and Infection Microbiology. Vol. 2 Article 53
  • Hoang, K.V., N. J. Stern, A. M. Saxton, F. Xu, X. Zeng, and J. Lin. 2011. Prevalence, development, and molecular mechanisms of bacteriocin resistance in Campylobacter. Applied and Environmental Microbiology 77:2309-2316
  • Zhou, W., and J. Lin. 2011. Functional cloning and characterization of antibiotic resistance genes from chicken gut microbiome. Submitted to Applied and Environmental Microbiology.
  • Zeng, X., F. Xu, Y. Mo, and J. Lin. 2011. A novel enterobactin esterase Cee plays a critical role in ferric enterobactin acquisition and pathogenesis of Campylobacter. Submitted to Proc. Natl. Acad. Sci.
  • Zeng, X., and J. Lin. 2011. Does Campylobacter use intracellular reductive iron release mechanism to utilize ferric enterobactin 92nd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 4-6, Chicago
  • Hoang, K., Y. Wang, and J. Lin. 2011. Identification of genetic loci required for Campylobacter resistance to fowlicidin-1, a chicken host defense peptide. Submitted to Frontiers in Cellular and Infection Microbiology
  • Hoang, K., and J. Lin. 2011. Identification and characterization of genes required for Campylobacter resistance to Fowlicidin-1, a chicken host defense peptide. 92nd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 4-6, Chicago
  • Zeng, X., F. Xu, and J. Lin. 2011. Complex molecular interaction of ferric enterobactin acquisition in campylobacter: role of TonB-ExbB-ExbB energy transduction system. 92nd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 4-6, Chicago
  • Mo, Y., X. Zeng, and J. Lin. 2011. Enzymatic characteristics of a novel enterobactin esterase Cee in Campylobacter. 92nd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 4-6, Chicago
  • Lin, J., X. Zeng, F. Xu, Y. Mo. 2011. Unique ferric enetrobactin iron acquisition system in Campylobacter. Fourth International Rushmore Conference on Enteric Diseases. Dec 3-4, Chicago.
  • Zeng, X., F. Xu, and J. Lin. 2011. Identification and characterization of a novel enterobactin esterase, Cee, in Campylobacter. 16th International Workshop on Campylobacter, Helicobacter and Related Organisms. Vancouver, Canada
  • Zeng, X., J. Lin. 2011. Identification of putative ferric reductase required for reductive iron release from ferric enterobactin in Campylobacter. 16th International Workshop on Campylobacter, Helicobacter and Related Organisms. Vancouver, Canada
  • Zeng, X., and J. Lin. 2011. The TonB-ExbB-ExbD energy transduction systems required for high-affinity ferric enterobactin acquisition in Campylobacter. 16th International Workshop on Campylobacter, Helicobacter and Related Organisms. Vancouver, Canada
  • Zhou, W., and J. Lin. 2011. Metagenomic discovery of novel antibiotic resistance genes in chicken gut microbiota and their functionality in Campylobacter. 16th International Workshop on Campylobacter, Helicobacter and Related Organisms. Vancouver, Canada